摘要
通过花粉管通道转基因技术,将E6启动子驱动的兔角蛋白基因导入高产棉花品种苏棉16号。所用转基因表达载体还含有选择标记基因NPTⅡ(卡那霉素抗性基因)及Gus报告基因。对转化体后代的检测结果表明,T1代有2.1%呈现Gus阳性,在Gus阳性株中84.6%具有卡那霉素抗性。用依据E6启动子序列和兔角蛋白基因序列设计的两对引物,对经过上述筛选的植株进行PCR检测,多次重复,最终确定3株结果稳定的转兔角蛋白基因棉株。从品质分析结果看,这3个株系成熟棉纤维的品质部分得到改良,尤其比强度有较大幅度提高,与转基因受体相比平均提高6.3cN·tex 1。
Cotton (Gossypium hirsutum L.) plants of the cultivar Sumian 16 were transformed with a rabbit keratin gene under the control of a fiber-specific E6 promoter, using pollen tube-mediated transformation. The plasmid used for the transformation also contained a Gus (β-glucuronidase) gene, and a NPTⅡ(neomycin phosphotransferase) gene driven by 35S promoters, respectively. After injection of the plasmid DNA into 500 ovaries of the cotton cultivar, 626 seeds were harvested. The seeds were then germinated on 0.7%(w/v) agar-solidified MS medium, and the seedlings were examined by a histochemical Gus assay and a kanamycin leaf-spotting test. The Gus assay revealed 13 positive seedlings, in which 11 were found kanamycin-resistance. To detect the presence of the keratin gene in the putatively transformed plants that survived the above selections, PCR analysis with keratin gene-specific primers was performed, and 3 plants showed an amplified product band of the expected size. The values of (2.5%) span length(SL), strength, and micronaire of the fiber were determined to evaluate the fiber quality of the rabbit keratin-transgenic cotton. The results indicated the fiber quality, to some extent, was improved, especially the average fiber strength was increased by 6.3 cN·tex^(-1)to that of Sumian 16.
出处
《棉花学报》
CSCD
北大核心
2004年第2期72-76,共5页
Cotton Science
基金
江苏省农业高技术项目(BG2002307)
